Preparation method and application of conditioned medium for deer periosteum
A culture medium and deer bone technology, applied in vertebrate cells, artificial cell constructs, animal cells, etc., can solve the problems of inability to provide a culture environment for tissues, inability to collect conditioned medium at any time, and difficulty in controlling the concentration of carbon dioxide. The effect of culture time, improving survival rate and convenient operation
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Embodiment 1
[0040] The invention provides a method for preparing conditioned medium by cultivating staghorn periosteum tissue in a triangular flask, the steps are as follows:
[0041] 1) Using animal tissue cutter ( figure 1 ) cutting the deer periosteum tissue collected from the living body into thin slices with a length of 6 mm, a width of 2 mm, and a thickness of 1.7 mm to obtain slices of the periosteum tissue;
[0042] 2) Slice 20 pieces of periosteum tissue obtained in step 1), and put them into a Erlenmeyer flask.
[0043] 3) Add nerve culture medium to the Erlenmeyer flask and fill it with saturated concentration of CO 2 Concentration, stopper the Erlenmeyer flask, put it in the incubator and cultivate it for 4 days, and collect the culture medium after the cultivation is completed;
[0044] 4) Filter the culture solution obtained in step 3) with a 0.22 μm filter membrane to obtain the velvet antler periosteum tissue-conditioned culture solution.
Embodiment 2
[0046] The invention provides a method for preparing the conditioned culture medium of periosteum tissue in deer antler area, the steps are as follows:
[0047] 1) Using an animal tissue cutter, cut the periosteum tissue from the deer antler area collected from the living body into thin slices with a length of 3 mm, a width of 2 mm, and a thickness of 0.7 mm to obtain slices of the periosteum tissue;
[0048] 2) Put 15 pieces of periosteum tissue slices obtained in step 1) evenly into an insert dish coated with rat tail collagen, and then put the insert dish into a 24-well culture plate ( figure 2 a) in the hole;
[0049] 3) Add nerve culture medium to the insert dish and the 24-well culture plate, that is, the upper chamber and the lower chamber of the culture, and store at 37°C, CO 2 Cultivate for 3 days under the condition of 5% concentration and saturated humidity, and collect the culture medium after the cultivation is completed;
[0050] 4) Filter the culture solution...
Embodiment 3
[0052] The invention provides a method for preparing conditioned culture medium of staghorn periosteum tissue, the steps are as follows:
[0053] 1) Use an animal tissue cutter to cut the periosteum tissue collected from the living body into thin slices with a length of 3 mm, a width of 2 mm, and a thickness of 0.7 mm to obtain periosteum tissue slices;
[0054] 2) Slice 50 pieces of periosteal tissue obtained in step 1), put them into an insert dish coated with rat tail collagen, and put the insert dish into the well of a 6-well culture plate;
[0055] 3) To the insert dish and 6-well culture plate ( figure 2 b) Add nerve culture medium, at 37°C, CO 2 Cultivate for 3 days under the condition of 5% concentration and saturated humidity, and collect the culture medium after the cultivation is completed;
[0056] 4) Filter the culture solution obtained in step 3) with a 0.22 μm filter membrane to obtain the velvet antler periosteum tissue-conditioned culture solution.
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Abstract
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